LOCUS       sequence-181455-        8181 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RcrCas13a for bacterial expression and contains backbone 
            for spacer cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8181)
  AUTHORS   Abudayyeh OO, Gootenberg JS, Essletzbichler P, Han S, Joung J, 
            Belanto JJ, Verdine V, Cox DBT, Kellner MJ, Regev A, Lander ES, 
            Voytas DF, Ting AY, Zhang F
  TITLE     RNA targeting with CRISPR-Cas13.
  JOURNAL   Nature. 2017 Oct 4. doi: 10.1038/nature24049.
  PUBMED    28976959
REFERENCE   2  (bases 1 to 8181)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2017 Oct 4. doi: 10.1038/nature24049."
COMMENT     Sequence Label: pC031 - RcrCas13a from Rhodobacter capsulatus ...
FEATURES             Location/Qualifiers
     source          1..8181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             59..1249
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(411..430)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(2283..2942)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2857..2876
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2943..3045)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3437..3454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3571..4116)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3651..3670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(4154..4159,4160..4177,4178..4183)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        8056..8090
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
ORIGIN
        1 caagctttaa tgcggtagtt tatcacagtt aaattgctaa cgcagtcagg caccgtgtat
       61 gaaatctaac aatgcgctca tcgtcatcct cggcaccgtc accctggatg ctgtaggcat
      121 aggcttggtt atgccggtac tgccgggcct cttgcgggat atcgtccatt ccgacagcat
      181 cgccagtcac tatggcgtgc tgctagcgct atatgcgttg atgcaatttc tatgcgcacc
      241 cgttctcgga gcactgtccg accgctttgg ccgccgccca gtcctgctcg cttcgctact
      301 tggagccact atcgactacg cgatcatggc gaccacaccc gtcctgtgga tcctctacgc
      361 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
      421 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
      481 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc
      541 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat
      601 gcaggagtcg cataagggag agcgtcgacc gatgcccttg agagccttca acccagtcag
      661 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
      721 catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt
      781 tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct
      841 cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat
      901 cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg
      961 gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca
     1021 ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag gatcgctcgc
     1081 ggctcttacc agcctaactt cgatcattgg accgctgatc gtcacggcga tttatgccgc
     1141 ctcggcgagc acatggaacg ggttggcatg gattgtaggc gccgccctat accttgtctg
     1201 cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgaa tggaagccgg
     1261 cggcacctcg ctaacggatt caccactcca agaattggag ccaatcaatt cttgcggaga
     1321 actgtgaatg cgcaaaccaa cccttggcag aacatatcca tcgcgtccgc catctccagc
     1381 agccgcacgc ggcgcatctc gggcagcgtt gggtcctggc cacgggtgcg catgatcgtg
     1441 ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa
     1501 tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca
     1561 acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcccct
     1621 acgtgctgct gaagttgccc gcaacagaga gtggaaccaa ccggtgatac cacgatacta
     1681 tgactgagag tcaacgccat gagcggcctc atttcttatt ctgagttaca acagtccgca
     1741 ccgctgccgg tagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
     1801 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgcccaac agtcccccgg
     1861 ccacggggcc tgccaccata cccacgccga aacaagcgcc ctgcaccatt atgttccgga
     1921 tctgcatcgc aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc
     1981 gctaaccgtt tttatcaggc tctgggaggc agaataaatg atcatatcgt caattattac
     2041 ctccacgggg agagcctgag caaactggcc tcaggcattt gagaagcaca cggtcacact
     2101 gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc
     2161 ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt catccgctta
     2221 ttatcactta ttcaggcgta gcaaccaggc gtttaagggc accaataact gccttaaaaa
     2281 aattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg
     2341 acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg
     2401 tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg
     2461 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgacac gaaaaacata
     2521 ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc
     2581 gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac
     2641 gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc
     2701 tcaccgtctt tcattgccat acggaattcc ggatgagcat tcatcaggcg ggcaagaatg
     2761 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta
     2821 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa
     2881 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc
     2941 attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat
     3001 cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcaacgtg tcattttcgc
     3061 caaaagttgg cccagggctt cccggtatca acagggacac caggatttat ttattctgcg
     3121 aagtgatctt ccgtcacagg tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac
     3181 ttactgattt agtgtatgat ggtgtttttg aggtgctcca gtggcttctg tttctatcag
     3241 ctgtccctcc tgttcagcta ctgacggggt ggtgcgtaac ggcaaaagca ccgccggaca
     3301 tcagcgctag cggagtgtat actggcttac tatgttggca ctgatgaggg tgtcagtgaa
     3361 gtgcttcatg tggcaggaga aaaaaggctg caccggtgcg tcagcagaat atgtgataca
     3421 ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag
     3481 cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca
     3541 gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca
     3601 tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca
     3661 ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg
     3721 tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt
     3781 aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg
     3841 ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg
     3901 cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc
     3961 taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa
     4021 gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca
     4081 gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata
     4141 aaatatttct agatttacac tttatgcttc cggctcgtat gttaggaggt cttatccatg
     4201 cagattggga aagtccaggg cagaaccatt agcgagtttg gcgaccccgc aggagggctg
     4261 aaaaggaaga ttagcaccga cggaaagaac cggaaagaac tgcccgccca tctgtcaagc
     4321 gatccaaagg ctctgatcgg ccagtggatt agcggcattg ataagatcta caggaagcct
     4381 gattcaagaa agagcgatgg aaaagccatc catagcccaa ctccaagcaa gatgcagttt
     4441 gatgccaggg acgacctggg ggaagcattc tggaaactgg tgtcagaggc tgggctggct
     4501 caggacagcg attacgatca gtttaaaagg cggctgcatc cctatgggga taagtttcag
     4561 cccgccgata gcggcgctaa actgaaattc gaagctgatc ctccagagcc ccaggctttt
     4621 cacggccgct ggtatggggc aatgagcaaa agaggaaacg acgctaaaga actggcagca
     4681 gcactgtatg aacatctgca tgtcgatgaa aaacgaattg atggccagcc aaaaaggaac
     4741 ccaaagacag ataaattcgc accagggctg gtcgtggcca gggcactggg gatcgagagc
     4801 tctgtgctgc cacgggggat ggcaagactg gcacggaatt ggggggaaga ggaaatccag
     4861 acatacttcg tggtcgacgt ggcagcaagc gtgaaggaag tggcaaaggc agccgtgagc
     4921 gccgcacagg ccttcgaccc acctcggcag gtgtccggca ggtcactgag cccaaaggtc
     4981 gggtttgccc tggctgaaca tctggagcgg gtcacaggct ccaaaaggtg cagcttcgat
     5041 cctgcagctg ggccaagcgt cctggccctg cacgacgaag tcaagaaaac ctataaacgg
     5101 ctgtgcgccc ggggaaagaa cgccgcaaga gcctttccag cagacaaaac agaactgctg
     5161 gccctgatgc ggcataccca cgagaaccgg gtgaggaatc agatggtcag aatggggcgg
     5221 gtgagtgagt atagggggca gcaggcaggg gacctggcac agagccacta ctggacaagc
     5281 gcaggacaga ccgaaatcaa ggagtcagag atttttgtcc gcctgtgggt cggggctttc
     5341 gcactggccg ggaggtcaat gaaagcctgg attgatccca tgggaaagat cgtcaacact
     5401 gaaaagaacg atcgcgacct gaccgctgca gtcaacattc ggcaggtcat cagcaacaag
     5461 gagatggtgg ccgaagctat ggcacgacgg gggatctact ttggagaaac cccagagctg
     5521 gataggctgg gagcagaagg caatgagggc ttcgtctttg cactgctgag atacctgaga
     5581 gggtgcagga atcagacctt tcacctgggg gccagggcag ggttcctgaa agagattaga
     5641 aaagaactgg aaaaaacccg gtgggggaag gccaaggagg cagagcacgt ggtcctgacc
     5701 gataaaactg tcgctgccat cagagccatc atcgacaacg atgccaaggc actgggggcc
     5761 cggctgctgg cagacctgtc aggggctttt gtcgctcact atgccagtaa agaacacttt
     5821 agcaccctgt actcagaaat tgtcaaggct gtgaaggacg caccagaggt gtcatccggg
     5881 ctgccccggc tgaagctgct gctgaagagg gcagacggag tcagaggcta tgtccatggg
     5941 ctgagggata caaggaaaca cgcctttgct actaaactgc caccacctcc agcaccaagg
     6001 gaactggatg acccagccac caaagccaga tacatcgcac tgctgagact gtacgatgga
     6061 ccattcaggg cttatgcatc cggcattact gggacagctc tggccgggcc agctgcacga
     6121 gccaaggagg ccgcaaccgc tctggcacag tccgtgaatg tcacaaaagc atacagcgat
     6181 gtgatggagg gcaggtcaag cagactgagg ccacccaacg atggggaaac actgagagaa
     6241 tacctgagcg cactgacagg agaaacagca acagaatttc gggtccagat tggctatgaa
     6301 agcgactcag agaatgctcg gaagcaggcc gagttcattg agaactatag acgggatatg
     6361 ctggctttta tgtttgagga ctacatccgg gctaaggggt tcgactggat tctgaaaatc
     6421 gagcccggag caactgcaat gacacgggca cctgtgctgc ccgagccaat tgacacccgg
     6481 gggcagtacg aacactggca ggcagcactg tatctggtaa tgcatttcgt gcccgcatca
     6541 gatgtcagca acctgctgca tcagctgagg aagtgggagg ccctgcaggg gaaatatgaa
     6601 ctggtgcagg atggcgacgc aacagatcag gcagacgcac gacgagaggc cctggatctg
     6661 gtcaagcgat ttcgcgatgt cctggtgctg ttcctgaaaa caggagaggc ccggttcgag
     6721 ggacgggcag ctccattcga cctgaaaccc ttccgagcac tgttcgcaaa tccagcaact
     6781 ttcgatcggc tgtttatggc tacacctacc acagcaaggc cagcagagga tgatcccgag
     6841 ggggacggcg catcagaacc agagctgagg gtggcccgga ctctgagagg gctgcggcag
     6901 attgccaggt ataatcacat ggcagtcctg agcgatctgt ttgcaaagca caaagtccga
     6961 gatgaggaag tcgcaaggct ggcagaaatt gaagatgaga ctcaggagaa atcacagatc
     7021 gtcgctgcac aggaactgag aaccgacctg catgacaaag tcatgaagtg tcatcccaaa
     7081 accatctctc ctgaagaaag acagtcctac gccgcagcta ttaagaccat cgaagagcat
     7141 aggtttctgg tggggagggt ctatctggga gatcatctgc ggctgcacag actgatgatg
     7201 gatgtgattg gacggctgat cgattacgct ggagcctacg agagggatac tgggacattt
     7261 ctgatcaatg cctcaaagca gctgggggcc ggcgcagact gggcagtgac aattgccggg
     7321 gcagcaaaca ctggcgccag gactcagact cggaaggacc tggcccactt taacgtgctg
     7381 gatagggccg acgggacacc tgacctgacc gccctggtga atcgagcaag agaaatgatg
     7441 gcatacgatc ggaaaagaaa aaacgcagtc ccaagatcaa ttctggatat gctggccagg
     7501 ctgggactga ctctgaaatg gcagatgaag gatcacctgc tgcaggatgc aactattacc
     7561 caggctgcaa ttaagcatct ggacaaagtg cggctgacag tgggcgggcc tgcagctgtg
     7621 actgaggcac gattctccca ggactacctg cagatggtgg ccgcagtgtt taacgggtct
     7681 gtccagaatc caaagccaag gaggagggac gacggcgacg catggcataa accacctaag
     7741 ccagcaactg cacagtccca gcccgaccag aagccaccca acaaagcacc tagcgcaggg
     7801 tcacggctgc ccccaccaca ggtcggggaa gtctatgaag gggtggtggt gaaagtgatc
     7861 gataccgggt cactgggatt cctggcagtc gaaggggtcg cagggaatat cggcctgcat
     7921 attagtaggc tgaggagaat cagagaagat gctattatcg tgggcaggag atacaggttt
     7981 tgggtcgaga tctacgtgcc ccccaagtct aatacctcca aactgaacgc tgctgacctg
     8041 gtgcgaattg actgattgac agctagctca gtcctaggta taatgctagc gcctcacatc
     8101 accgccaaga cgacggcgga ctgaacggag acggagattc gtctcggcct cacatcaccg
     8161 ccaagacgac ggcggactga a
//
